Summary of Toxicology Data - · PDF fileSUMMARY OF TOXICOLOGY DATA FLUXAPYROXAD ... (DPN) # 53118 SB 950 # (Not applicable) Original ... purity 99.7%, for 2 yrs in an oncogenicity

CALIFORNIA ENVIRONMENTAL PROTECTION AGENCYDEPARTMENT OF PESTICIDE REGULATION

MEDICAL TOXICOLOGY BRANCH

SUMMARY OF TOXICOLOGY DATAFLUXAPYROXAD

Chemical Code # 6033, Document Processing Number (DPN) # 53118

SB 950 # (Not applicable)Original date: Oct 10, 2011

Revised date: (Not applicable)

I. DATA GAP STATUS Chronic toxicity, rat: No data gap, no adverse effect (but see oncogenicity, rat)

Chronic toxicity, dog: No data gap, possible adverse effect

Oncogenicity, rat: No data gap, possible adverse effect

Oncogenicity, mouse: No data gap, no adverse effect

Reproduction, rat: No data gap, no adverse effect

Developmental toxicity, rat: No data gap, no adverse effect

Developmental toxicity, rabbit: No data gap, no adverse effect

Gene mutation: No data gap, no adverse effect

Chromosome effects: No data gap, possible adverse effect

DNA damage: No data gap, no adverse effect

Neurotoxicity: Not required at this time

Toxicology one-liners are attached. All record numbers for the above study types through 254228 (Document No. 53118-0083) wereexamined. This includes all relevant studies indexed by DPR as of 8/29/11.

In the 1-liners below: ** indicates an acceptable study. Bold face indicates a possible adverse effect. ## indicates a study on file but not yet reviewed.

File name: t20111010.wpdRevised by: C. Aldous, 10/10/11(original)

DPR MEDICAL TOXICOLOGY FLUXAPYROXAD TOX SUMMARY t20111010.wpd Page 2

TABLE OF CONTENTS

COMBINED, RAT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3

CHRONIC TOXICITY, DOG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

ONCOGENICITY, MOUSE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4

REPRODUCTION, RAT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

DEVELOPMENTAL TOXICITY, RAT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

DEVELOPMENTAL TOXICITY, RABBIT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5

GENE MUTATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 6

CHROMOSOME EFFECTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

DNA DAMAGE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

NEUROTOXICITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

METABOLISM . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

SUBCHRONIC (and subacute, if applicable) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11DOG . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11RAT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12MICE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

MECHANISTIC STUDIES RELATING TO COMBINED, RAT . . . . . . . . . . . . . . . . . . . . . . 13

IMMUNOTOXICITY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

TOXICITY STUDIES ON TECHNICAL IMPURITIES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

TOXICITY STUDIES ON METABOLITES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19BASF Reg. No. 5435595 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19BASF Reg. No. 5069089 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20BASF Reg. No. 5621781 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22BASF Reg. No. 5570265 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22


II. TOXICOLOGY ONE-LINERS AND CONCLUSIONS These pages contain summaries only. Individual worksheets may identify additional effects.

COMBINED, RAT

**53118-0051 254137 Buesen, R., V. Strauss, A. Groeters, E. Fabian, and W. Mellert, “BAS700 F - Combined chronic toxicity/carcinogenicity study in Wistar rats - administration via thediet up to 24 months,” BASF, Ludwigshafen, Germany, 10/19/09. Laboratory Study #2009/1072490. Groups of fifty rats/sex/group were dosed in diet with BAS 700 F[Fluxapyroxad], purity 99.7%, for 2 yrs in an oncogenicity study at 0, 50, 250, 1500, or 3000ppm. An additional 10/sex/group were dosed at those levels for 12 months as satellite chronicstudy rats. Estimated mean lifetime study exposures were 2.1, 11, 68, and 145 mg/kg/day forincreasing dose groups of males, and 2.7, 14, 82, and 182 mg/kg/day for females. NOEL = 50ppm [M: 2.1 mg/kg/day, F: 2.7 mg/kg/day], with liver as primary target organ. Centrilobularhypertrophy was elevated in lifetime study at 250-3000 ppm in both sexes, in satellite studymales at 1500-3000 ppm and females at 250-3000 ppm. Liver absolute and relative weightswere elevated at 250-3000 ppm in satellite study females and in males in the lifetime study, andat 1500-3000 ppm in lifetime study females and satellite study males. The relative liver weightsof the 250 ppm lifetime study females were also elevated. The most consistent clinicalchemistry change was dose-related increase in cholesterol at 250-3000 ppm in males and at1500-3000 ppm in females. Various other findings at 250 ppm and above included decreasedbody weight in females (final yr of study) and a sharply dose-related increase in ferric irondeposition in femur cancellous bone evidenced by Perl’s Prussian Blue stain in both sexes. There was a dose-related increase in hepatocellular adenoma in 1500-3000 ppm males and in3000 ppm females. Hepatocellular carcinoma was elevated significantly in 3000 ppm males. Non-significant increases in hepatocellular adenomas in 250 ppm males and in 1500 ppmfemales were plausibly also treatment-related. Additional associated hepatotoxicity wasmanifested by strongly dose-related spongiosis in 1500-3000 males, and by pigmentaccumulation (probably lipofuscin) in 1500-3000 ppm males and females. A strong hyperostosisresponse was observed in bones of the skull in 3000 ppm males and females. Thyroid glandfollicular cell hyperplasia was observed in 1500-3000 ppm males. Tooth whitening wasobserved in incisors of many males and females at 1500-3000 ppm, without evident microscopicchange. Thyroid follicular cell hyperplasia was elevated in 1500-3000 ppm males. Thyroidfollicular cell combined adenoma plus carcinoma incidence was statistically elevated in 3000ppm males compared to concurrent controls. Overall follicular cell tumor incidence was withinhistorical control range, yet some supplementary studies cited in the present DPR review supportthe plausibility of a treatment effect on follicular tumors, likely mediated by enhanced livermetabolism of thyroxin. Study is acceptable. Hepatocellular tumors are “possible adverseeffects:” noted to exist along with predisposing pathology. Aldous, 8/31/11.

CHRONIC TOXICITY, RAT

See COMBINED, RAT (above)


CHRONIC TOXICITY, DOG

**53118-0049 254135 Hempel, K., V. Strauss, S. Groeters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Chronic toxicity study in beagle dogs - administration in the diet for 12 months,”BASF SE, Ludwigshafen, Germany, 10/15/09. Laboratory Study # 2008/1090458, Report No.33D0683/05088. Groups of 5 beagles/sex/group were dosed in diet with BAS 700 F(fluxapyroxad, Batch COD-001026, purity 99.4%) for 1 year in a chronic study. Dietary levelswere 0, 300, 1500, and 12000 ppm for males, and 0, 300, 1500, and 9000 ppm for females. Achieved dose levels for respective male treated groups were 8, 39, and 335 mg/kg/day. Corresponding levels for females were 9, 43, and 257 mg/kg/day. NOEL = 300 ppm, based ondose-related iron staining (Perl’s Prussian blue positive) in liver and spleen (both sexes), andfibrosis in liver (females). Livers showed fine granular intra-cytoplasmic staining, mostpronounced in the peri-portal areas, largely in subcapsular locations. Splenic pigmentconcentrated in intra-cytoplasmic areas of connective tissue elements, and was evidently nothemosiderin. Liver-associated clinical chemistry changes included dose-related decreases incirculating albumin with a probable secondary reduction in circulating calcium in males, reducedbilirubin (both sexes), and reduced cholesterol (males). Gallbladders had pigmentation in bothsexes at 1500 ppm and above. All of the above findings were evident in both sexes at respectivehighest dose levels. Other findings at 9000-12000 ppm included marked reduction of foodconsumption in females and prostate atrophy in males (reduced tissue mass, but normalmicroscopic appearance). The following were seen in both sexes at 9000-12000 ppm: transitoryvomiting (at initiation of study), modest body weight decrements, a general increase in plateletcounts, very sharply increased serum alkaline phosphatase and gamma-glutamyltransferase(consistent with hepato-biliary toxicity), and sharply elevated liver weights. Study is acceptable,with liver fibrosis as a “possible adverse effect.” Aldous, Oct. 7, 2011.

ONCOGENICITY, RAT

See COMBINED, RAT (above)

ONCOGENICITY, MOUSE

**53118-0050 254136 Buesen, R., K. Kuettler, V. Strauss, and B. van Ravenzwaay, “BAS 700F - Carcinogenicity study in C57BL/6 J Rj mice - administration via the diet over 18 months”(including Amendment No. 1), BASF SE, Ludwigshafen, Germany, Feb. 4, 2010 (amended). BASF Registration Document No. 2010/7003500. Report No. 87C0683/05082. Groups of 50mice/sex/group were dosed in diet with 0, 150, 750, 3000, or 6000 ppm fluxapyroxad (BatchCOD-001049, purity 99.2%) in the oncogenicity study (18 months). An additional 10/sex wereassigned to satellite groups at 0 or 6000 ppm (9 months). Estimated achieved dose levels intreated oncogenicity study males were 21, 107, 468, and 996 mg/kg/day, respectively. Corresponding female dose levels were 33, 158, 652, and 1307 mg/kg/day. NOEL = 150 ppm,based on macrovesicular fatty change in livers of both sexes at 750 ppm and above inoncogenicity study mice. Treatment-related increases in liver weights were observed in males at750 ppm and above, and in females at 3000 and 6000 ppm. Body weights were reduced in dose-related fashion in 3000 to 6000 ppm males, and transiently also in 6000 ppm females. Incisors,


particularly mandibular, had a whitish discoloration at 3000-6000 ppm in both sexes, with 2affected females at 750 ppm. Study is acceptable, with no adverse effects. Aldous Oct. 6, 2011.

REPRODUCTION, RAT

**53118-0054 254140 Schneider, S., V. Strauss, S. Groeters, and W. Mellert, “BAS 700 F -Two-generation reproduction toxicity study in Wistar rats - administration via the diet,” BASFSE, Ludwigshafen, Germany, 10/21/09. Laboratory Study # 2009/1072491, Report No.70R0683/05092. Wistar rats, 25/sex/group, were dosed in diet to achieve 0, 10, 50, or 300mg/kg/day in a standard 2-generation reproduction study. Parental systemic toxicity NOEL < 10mg/kg/day, based on increased absolute liver weights in F1 adults, and dose-related centrilobularhypertrophy in both sexes of both generations. Parental reproductive effects NOEL = 300mg/kg/day (highest dose tested). Offspring viability and growth NOEL = 50 mg/kg/day (bodyweight decrement for pups in both generations). The highest dose level was clearly rigorous,eliciting significant decrements in food consumption and body weight, and additional signs ofmarked liver toxicity at that dose included gross enlargement and discoloration, hepatocellularnecrosis in 5-6 males/generation, and highly elevated serum gamma-glutamyltransferase (SGGT)in both generations of both sexes. Thyroid follicular hypertrophy/hyperplasia and altered colloidwere observed at 50-300 mg/kg/day. Acceptable, with no adverse effects. Aldous, Oct. 6, 2011.

DEVELOPMENTAL TOXICITY, RAT

**53118-0052 254138 Buesen, R., V. Strauss, W. Kaufmann, E. Fabian, and B. vanRavenzwaay, “BAS 700 F - Prenatal developmental toxicity study in Wistar rats - oral administration (gavage),” BASF SE, Ludwigshafen, Germany, Oct. 8, 2009. Laboratory Study #2009/1072492, Project No. 30R0683/05094. Groups of 25 mated female Wistar rats/group weredosed with BAS 700 F (fluxapyroxad, Batch COD-000899, purity 99.7%) daily from gestationdays 6-19 at 0, 25, 200, or 1000 mg/kg/day. Maternal NOEL < 25 mg/kg/day, based on modestincreases in relative thyroid gland weights. Developmental toxicity NOEL = 1000 mg/kg/day(no effect at highest dose tested). Common dose-related maternal changes at 200 to 1000mg/kg/day included transiently reduced food consumption during the first 2 days of dosing, withsignificant body weight gain decrements during that period. Reduced serum bilirubin andincreased albumin, each likely related to altered liver function, were observed in the same doserange. Study is acceptable, with no adverse effects. Aldous, Oct. 6, 2011.

DEVELOPMENTAL TOXICITY, RABBIT

**53118-0053 254139 Buesen, R., E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Prenataldevelopmental toxicity study in Himalayan rabbits - oral administration (gavage),” BASF SE,Ludwigshafen, Germany, Oct. 8, 2009, Laboratory Study # 2009/1072493, Project No.40R0683/05089. Groups of 25 artificially inseminated rabbits/group were dosed with BAS 700F (fluxapyroxad, Batch COD-000899, purity 99.7%) daily from gestation days 6-28, at 0, 10, 25,or 60 mg/kg/day. Maternal NOEL = 25 mg/kg/day. Food consumption was sharply reduced inhigh dose does through about day 22. Body weight gain was reduced in these does between days


9 and 16. Additionally, clinical signs of “reduced defecation” were noted in 8 high dose does. Developmental NOEL = 25 mg/kg/day (paw hyperflexion). Study is acceptable, with no adverseeffects. Aldous, Oct. 6, 2011.

GENE MUTATION

**53118-0055 254141 Schulz, M. and R. Landsiedel, “BAS 700 F - Salmonella typhimurium /Escherichia coli reverse mutation assay (standard plate test and preincubation test),” BASF SE,Ludwigshafen, Germany, 10/13/08, Laboratory Study # 2008/1028479. This standard reversemutation assay used three replicates/dose with and without S-9 activation. Salmonella strainswere TA 98, TA 100, TA 1535, and TA 1537. E. coli strain was WP2 uvrA. Dose levels forfluxapyroxad [BAS 700 F (Batch COD-001026, purity 99.4%)] were uniformly 0, 20, 100, 500,2500, and 5000 :g/plate. There were no remarkable increases in revertants. Precipitation wasobserved in all groups at 500 :g/plate and above. Commonly there were dose-related reductionsin the numbers of revertants at 2500 to 5000 :g/plate, indicating cytotoxicity. Titer of viablecells, assessed in 2500 to 5000 :g/plate groups with S-9 activation, was commonly reduced indose-related fashion in the latter dose range. Positive controls were functional. Study isacceptable, with no adverse effects. Aldous, Oct. 4, 2011.

**53118-0055 254142 Schulz, M. and R. Landsiedel, “BAS 700 F: Salmonella typhimurium /Escherichia coli reverse mutation assay (standard plate test and preincubation test),” Oct. 6,2009, Laboratory Study # 2009/1080768. This standard reverse mutation assay used threereplicates/dose with and without S-9 activation. Study is analogous to Record No. 254141, butused a different batch [No. 35575/18] of BAS 700 F [Fluxapyroxad], purity 96.7%. Salmonellastrains were TA 98, TA 100, TA 1535, and TA 1537. E. coli strain was WP2 uvrA. Dose levelsfor fluxapyroxad were normally 0, 21, 106, 2650, and 5300 :g/plate (maximum dose levels were2650 and 530 :g/plate, respectively, for TA 1537 and TA 98 in preincubation assay only). There were no increases in revertants. Precipitation was observed in all groups at 530 :g/plateand above. Commonly there were dose-related reductions in revertants at 2650 to 5300 :g/plate,indicating cytotoxicity. Titer of viable cells, assessed in the two highest two dose groups with S-9 activation, was commonly reduced in dose-related fashion in the latter dose range, and possiblyas low as 265 :g/plate in the preincubation test of TA 98 with activation. Positive controls werefunctional. Study is acceptable, with no adverse effects. Aldous, Oct. 4, 2011.

**53118-0055 254144 Schulz, M. and R. Landsiedel, “In vitro gene mutation test in CHO cells(HPRT locus assay) with Reg. No. 5094351,” BASF AG, Ludwigshafen, Germany, 6/19/07. Laboratory Study # 2007/1020715. Test article was BAS 700 F [Fluxapyroxad], Batch COD-000899, purity 99.7%. There were two experiments, each with and without S-9. Limitingcytotoxicity ranged from 50 to 75 :g/ml. The first experiment used dose levels of 0, 5, 10, 20,50, and 100 :g/ml fluxapyroxad. The second experiment used 6.3, 12.5, 25, 50, 75, and 100:g/ml. There were two flasks per dose level. About 1 x 106 cells per flask were initiated onstudy. Treatment did not elicit mutagenicity. Positive controls (EMS without S-9 and MCAwith S-9) were functional. Acceptable. No adverse effects. Aldous, 6/29/11.

**53118-0055 254146 Schulz, M. and R. Landsiedel, “BAS 700 F: In vitro gene mutation testin CHO cells (HPRT locus assay),” BASF SE, Ludwigshafen, Germany, Oct. 2, 2009,2009/1078663. Test article was BAS 700 F [Fluxapyroxad], Batch 35575/18, purity 96.7%.


There were two initial experiments, each with and without S-9. A third experiment with S-9 wasundertaken for technical reasons. Limiting cytotoxicity was 75 :g/ml without S-9, and about120-125 :g/ml with S-9. There were two flasks per dose level. About 1 x 106 cells per flaskwere initiated on study. Treatment did not elicit mutagenicity. Positive controls (EMS withoutS-9 and MCA with S-9) were functional. Acceptable: there were sufficient concentrationsclosely spaced near to optimal levels for a thorough assessment. No adverse effects. Aldous,6/29/11.

CHROMOSOME EFFECTS

**53118-0056 254156 Schulz, M. and R. Landsiedel, “Cytogenetic study in vivo with LS5094351 in the mouse micronucleus test after two oral administrations,” BASF AG,Ludwigshafen, Germany, Dec. 6, 2006. Laboratory Study # 2006/1032708. Five maleCrl:NMRI mice/group were dosed twice orally (10 ml/kg corn oil suspension) with 0, 500, 1000,or 2000 mg/kg LS 5094351 (fluxapyroxad, Batch COD-000826, purity 99.6%) 24 hrs beforesacrifice. Positive controls were 20 mg/kg cyclophosphamide and 0.15 mg/kg vincristine sulfate,each with 24-hr intervals. No treatments caused clinical effects. Fluxapyroxad groups had noincreases in micronuclei. Positive controls were functional, with cyclophosphamide elicitingnumerous small micronuclei, and vincristine causing large numbers of large and smallmicronuclei. Acceptable, with no adverse effects. Aldous, Oct. 5, 2011.

**53118-0057 254157 Schulz, M. and R. Landsiedel, “BAS 700 F - Micronucleus test in bonemarrow cells of the mouse,” BASF SE, Ludwigshafen, Germany, Oct. 2, 2009. LaboratoryStudy # 2009/1072522. Five male Crl:NMRI mice/group were dosed ip (in 4 ml/kg DMSO)with 0, 500, 1000, or 2000 mg/kg BAS 700 F [Fluxapyroxad, Batch 35575/18, purity 96.7%], 24hrs before sacrifice. An additional 5 males/group received 0 or 2000 mg/kg fluxapyroxad, with a48-hr interval to sacrifice. Positive controls were 20 mg/kg cyclophosphamide and 0.15 mg/kgvincristine sulfate, each with 24-hr intervals. No treatments caused clinical effects. Fluxapyroxad groups had no increases in micronuclei. Positive controls were functional, withcyclophosphamide eliciting numerous small micronuclei, and vincristine causing large numbersof large and small micronuclei. Acceptable, with no adverse effects. Aldous, Oct. 6, 2011.

**53118-0056 254148 Schulz, M. and R. Landsiedel, “BAS 700 F - In vitro chromosomalaberration assay in V79 cells,” BASF AG, Ludwigshafen, Germany, 1/22/08. Laboratory Study# 2007/1023153. V79 cell line cultures of about 30000 to 80000 cells per culture were treatedwith at least 0, 12.5, 25, 50, and 100 :g/ml BAS 700 F [Fluxapyroxad], Batch COD-000899,purity 99.7% in two experiments, with 2 cultures per treatment combination (from which 200spreads were evaluated per treatment combination). Exposure durations and total time to harvestin the first experiment were 4 hrs/18 hrs with or without S-9. In the second experiment, thesetimes were 18 hrs/18 hrs and 18 hrs/28 hrs without S-9, and 4 hrs/28 hrs with S-9. Colcemid wasused to arrest mitosis. In all cases, 50 :g/ml was the maximum dose with sufficient readablespreads for analysis. Fluxapyroxad did not elicit chromosomal aberrations. Mitotic index wasunaffected by treatment. Study is acceptable, with no adverse effects. Aldous, 6/30/11.

**53118-0056 254149 Schulz, M. and R. Landsiedel,“BAS 700 F: In vitro chromosomalaberration assay in V79 cells,” BASF SE, Ludwigshafen, Germany, Oct. 1, 2009. Laboratory


Study # 2009/1078662. V79 cell line cultures of about 30000 to 80000 cells per culture weretreated with BAS 700 F [Fluxapyroxad], Batch 35575/18, purity 96.7%, in 3 experiments, with 2cultures per treatment combination (from which normally 200 spreads were evaluated pertreatment combination). Exposure durations and total time to harvest in the first and thirdexperiments were 4 hrs/18 hrs with or without S-9. In the second experiment, these times were18 hrs/18 hrs and 18 hrs/28 hrs without S-9, and 4 hrs/28 hrs with S-9. Colcemid was used toarrest mitosis. Fluxapyroxad elicited chromosomal aberrations in the entire range of 60 to 80:g/ml Experiment 3 (significantly elevated compared to concurrent controls, and outside thehistorical range at all three dose levels, with and without S-9). This range was close to toxicitylimits, with cell numbers at 80 :g/ml reduced to 59% of controls without S-9, and to 48% ofcontrols with S-9. Also, in the case of treatment without S-9 in Experiment 3, cell morphologywas atypical (“rounded”) and cells had markedly reduced attachment compared to controls at 40-80 :g/ml. In contrast, these treatment levels in groups treated with S-9 in Experiment 3 hadnormal cell morphology. In Experiment 1, there were small increases in chromosomalaberrations at the maximum readable dose of 62.5 :g/ml, with and without S-9 (not statisticallysignificant compared to concurrent controls, but outside historical control range for thelaboratory). Incidence of polyploidy was unaffected by treatment. Study is acceptable, with a“possible adverse effect” of chromosomal aberrations. Aldous, 6/30/11.

DNA DAMAGE

**53118-0057 254158 Schulz, M. and R. Landsiedel, “BAS 700 F - In vivo unscheduled DNAsynthesis (UDS) assay in rat hepatocytes,” BASF AG, Ludwigshafen, Germany, Nov. 8, 2008(amended). Laboratory Study # 2008/7020135. Groups of 3 male Wistar rats/group were dosedby oral gavage with 0, 1000, or 2000 mg/kg BAS 700 F [Fluxapyroxad], Batch COD-000899,purity 99.7%, or with 50 mg/kg 2-acetylaminofluorene (positive control) with sampling times of3 hrs or 14 hrs to sacrifice. Isolated hepatocytes were allowed to attach to wells, treated with 3H-thymidine for 4 hrs, then washed and incubated without radiolabel for another 12 hrs. Cells wereprocessed with photographic emulsions in darkness to visualize 3H disintegration tracks, andstained with H&E. Coded slides were examined for net (nuclear minus cytoplasmic) graincount, with proliferating cells censored. All 2000 mg/kg fluxapyroxad rats showed clinical signsat sacrifice (apathy). Fluxapyroxad did not elicit UDS, whereas positive controls werefunctional. Study is acceptable, with no adverse effects. Aldous, Oct. 6, 2011.

**53118-0057 254159 Schulz, M. and R. Landsiedel, “BAS 700 F: In vivo unscheduled DNAsynthesis (UDS) assay in rat hepatocytes,” BASF AG, Ludwigshafen, Germany, Oct. 1, 2009. Laboratory Study # 2009/1078661. Groups of 3 male Wistar rats/group were dosed iv with 0,2.5 or 5.0 mg/kg BAS 700 F [Fluxapyroxad], Batch 35575/18, purity 96.7%, or by gavage with50 mg/kg 2-acetylaminofluorene (positive control) with sampling times of 3 hrs or 14 hrs tosacrifice. Isolated hepatocytes were allowed to attach to wells, treated with 3H-thymidine for 4hrs, then washed and incubated without radiolabel for another 12 hrs. Cells were processed withphotographic emulsions in darkness to visualize 3H disintegration tracks, and stained with H&E. Coded slides were examined for net (nuclear minus cytoplasmic) grain count, with proliferatingcells censored. There was a dose-response for clinical signs in 2.5 to 5.0 mg/kg fluxapyroxadrats. Piloerection was observed at sacrifice of all fluxapyroxad rats. Fluxapyroxad did not elicitUDS, whereas positive controls were functional. Study is acceptable, with no adverse effects. Aldous, Oct. 6, 2011.


NEUROTOXICITY

53118-0040 254126 Kaspers, U., W. Kaufmann, E. Fabian, and B. van Ravenzwaay, “BAS 700F - Acute oral neurotoxicity study in Wistar rats - administration via gavage,” BASF SE,Ludwigshafen, Germany, Oct. 7, 2009. Laboratory Study # 2009/1065774, Project No.61S0683/05102. Groups of 10/sex/group were dosed once by gavage (in 1% CMC, 10 ml/kg) at0, 125, 500, or 2000 mg/kg. FOB and motor activity were assessed pre-study (Day -7), Day 0(after dosing), Day 7, and Day 14 on all rats. Five/sex were examined in controls and at 2000mg/kg at termination for neurohistopathology. NOEL = 125 mg/kg, with diminished motoractivity at Day 0 as the major finding in both sexes at 500 to 2000 mg/kg. Other findings, bothat Day 0, were significantly decreased rearing behavior in 500 to 2000 mg/kg males, andincreased landing foot splay in 2000 mg/kg males. Study is not acceptable, but upgradeable:validation of laboratory technicians’ ability to identify neurotoxic endpoints is requested. Thisusually entails positive control validation studies: such studies referenced in the present reportwere not current, but rather were completed about 7 years or more before this study wasundertaken. Aldous, Oct. 7, 2011.

53118-0048 254134 Kaspers, U., V. Strauss, W. Kaufmann, E. Fabian, and W. Mellert, “BAS700 F - Repeated dose 90-day oral neurotoxicity study in Wistar rats; administration in the diet,”BASF SE, Ludwigshafen, Germany, Nov. 2, 2009 (amended), Laboratory Study #2009/7006263, Project No. 63S0683/05090. Groups of 10 Wistar [Crl:WI(Han)] rats/sex/groupwere dosed in diet with BAS 700 F (fluxapyroxad, Batch COD-001026, purity 99.4%) at 0, 200,1000, or 5000 ppm for 3 months. Mean achieved dose levels were 11.5, 58, and 302 mg/kg/dayfor males, and 13.4, 67, and 338 mg/kg/day for females. Apparent NOEL for neurotoxicity =5000 ppm (highest dose tested). NOEL for general toxicity < 200 ppm, due primarily to liverand thyroid changes. Hepatocellular hypertrophy was observed in all treated males and in allfemales from 1000 to 5000 ppm: in each case there was a clear dose-response in degree ofhypertrophy. Thyroid weights were progressively elevated in all treatment groups of both sexes. Study protocol did not require microscopic examination of thyroid glands. Major clinicalchemistry changes, plausibly related to liver toxicity or to altered liver function, includedelevated globulins (all treated male groups, and 1000-5000 ppm females) and significantlyelevated albumin at 5000 ppm in both sexes. Serum inorganic phosphorus and calcium wereelevated at 5000 ppm in both sexes (possibly reflecting elevated serum carrier proteins). Glucose was sharply reduced in 5000 ppm males. Triglycerides were markedly increased at5000 ppm, particularly in females. Serum GGT was increased at 5000 ppm in both sexes:profoundly so in males. Study is not acceptable, but upgradeable: validation of laboratorytechnicians’ ability to identify neurotoxic endpoints is requested. This is usually done bypositive control validation studies. Such studies referenced in the present report were notcurrent: they were completed about 7 years or more before this study was undertaken. Noadverse effects. Aldous, Oct. 7, 2011.

METABOLISM

The following two studies fulfill the biokinetics/metabolic disposition data requirementsfor fluxapyroxad.


53118-0057 254160 Fabian, E. and R. Landsiedel, “14C-BAS 700 F: Study on the biokineticsin rats,” BASF SE, Ludwigshafen, Germany, 9/30/09. Laboratory Study # 2009/1074879. Parent is 3-(difluoromethyl)-1-methyl-N-(3',4',5'-trifluoro[1,1'-biphenyl]-2-yl)1H-pyrazole-4-carboxamide. Unlabeled fluxapyroxad was Batch No. COD-000842, 99.2% purity. Pyrazole-4-14C label (Batch No. 900-1101, radiochemical purity 99.8%, estimated chemical purity is 99%)was used for blood and plasma kinetics at 5, 50, and 500 mg/kg. Aniline-U-14C label, Batch No.916-1018, radiochemical purity 98.2%, chemical purity 97.3%, was used at 7.5 and 150 mg/kgfor balance and excretion studies, tissue distribution, and biliary studies. Examination of exhaledair for 14C-CO2 residues found negligible label. Record No. 254161 found minimal cleavagebetween pyrazole and aniline label positions, so that the label placement was arbitrary: anilinelabel was usually used for the present study. Usually 4 Wistar rats/sex were used at each studyphase, dose, and time point combination. At 7.5 mg/kg, about 85-90% of administered dose wasfound in feces, with 10% and 17% of administered dose in urine of males and females,respectively. Percent of label in urine after 150 mg/kg was reduced in both sexes. About 51-63% of administered dose was excreted via bile in cannulated rats of either sex at 7.5 and 150mg/kg, thus absorbed dose was about 60-70% of administered dose. Tissue levels after 7 dayswere low: liver at about 0.1% of administered dose being the most noteworthy. Peak plasmaconcentrations were delayed as dose increased. Tmax was 1, 8, and 24 hrs for 5, 50, and 500mg/kg, regardless of sex. Initial t1/2 in plasma ranged from 12-15 hr for 5 mg/kg to 25-27 hrs for500 mg/kg. Terminal t1/2 was > 30 hrs in all cases. Tissue levels in 7.5 mg/kg rats declinednormally over time. Highest initial levels (excluding alimentary tract) were adrenal glands,followed in order by liver and thyroid. In females, adipose tissue had higher remaining specificactivity than any of these organs at 48 hrs. The 150 mg/kg rats did not indicate remarkableaffinity for any tissue, although liver and adipose tissue levels were the highest 16-hr levels. Bythe time of the final assessment (96 hrs for males and 104 hrs for females), liver had the highestconcentration of assessed tissues. Useful segment of the metabolism study series. Aldous, Oct.6, 2011.

53118-0058 254161 Schopfer, C. and S. Labib, “The metabolism of 14C-BAS 700 F (Reg. No:5094351) in Wistar rats,” BASF SE, Limburgerhof, Germany, 9/24/09. Laboratory Study #2009/1019789. Samples were primarily taken from the in-life study, 53118-0057 254160 (seeSummary of Toxicology Data). Record No. 254160 reported that about 85-90% of administereddose was found in feces. Major metabolites were hydroxylation products of phenyl rings orconjugates thereof. Over one-half of labeled content also had lost the methyl group of thepyrazole. The present study reported disposition of low and high doses of fluxapyroxad (7.5 and150 mg/kg, respectively). Findings at the 7.5 mg/kg best represent plausible human exposures,and are emphasized in this review. The most abundant fecal metabolite in 7.5 mg/kg males wasM700F009 (product of N-demethylation of the pyrazole group, and of hydroxylation of one ofthe phenyl groups). This was also the dominant metabolite in 7.5 mg/kg females. M700F009 constituted 22% of administered dose in fecal extracts of males, and 53% in females. Metabolite M700F005 (parent with hydroxylation of one of the phenyl groups) constituted 9% ofadministered dose in feces of males and females. Biliary label accounted for 54-69% ofadministered dose in cannulated rats, with little influence of sex or dose. The most abundantmetabolite in bile of 7.5 mg/kg males (14% of administered dose) was M700F004, a glucuronideof phenyl-hydroxylated parent. M700F004 was also an important biliary metabolite in females(11% of administered dose). Conjugated (glucuronide, cysteine) products of M700F009 and ofM700F005 (and of isomers of these two metabolites) were quite common in the bile. The


comparatively large amount of M700F004 in bile compared to feces or urine, plus the moreplentiful M700F009 and other conjugated metabolites in feces, gives the impression thatintestinal microflora have appreciable glucuronidase activity, as well as possibly N-demethylaseactivity. Parent fluxapyroxad constituted about 3% of administered dose in feces of either sex. Parent was not found in urine nor bile. In contrast to the above, the most striking feature of highdose treatment was reduced absorption: parent fluxapyroxad constituted 44% of administereddose in males and 34% of administered dose in the feces of the 150 mg/kg males and females,respectively. At the plasma Tmax for low dose levels (1 hr), liver label concentration was higherthan other sampled tissues. In these low dose rats at Tmax, parent constituted 3.0% and 3.7% ofadministered dose in the livers of males and females, respectively. Parent comprised 59-70% oflabel in liver at that time. Useful data in support of metabolism data requirements. Aldous, Oct.4, 2011.

SUBCHRONIC (and subacute, if applicable)

DOG

**53118-0046 254132 Hempel, K., V. Strauss, A. Gröters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Repeated dose 90-day oral toxicity study in beagle dogs - administration in thediet,” BASF SE, Ludwigshafen, Germany, 7/16/09. Laboratory Study # 2008/1013661, ReportNo. 31D0683/05084. Groups of 5 beagles/sex/group were dosed in diet with BAS 700 F(fluxapyroxad, Batch COD-000899, purity 99.7%) for 90 days in a subchronic study. Dietarylevels were 0, 300, 1500, and 10000 ppm for males, and 0, 300, 1500, and 7500 ppm for females. Achieved dose levels for respective male treated groups were 9, 45, and 295 mg/kg/day. Corresponding levels for females were 10, 51, and 238 mg/kg/day. NOEL = 300 ppm in bothsexes, based on very slight, dose-related reduction of serum albumin. Other possible hepato-biliary responses were elevated alkaline phosphatase and serum (-glutamyltransferase at 7500and 10000 ppm. Also at these doses, levels of circulating calcium (which is largely transportedon albumin) were measurably reduced. A reduction in total bilirubin at 7500-10000 ppm likelyalso reflected altered liver function. Liver weights were significantly elevated at these levels,without associated histopathology. Food consumption was reduced in 7500 ppm females. Allhigh dose dogs vomited on the first day, sometimes up to 2 days: an apparent acute effect. Acceptable, with no adverse effects. Aldous, Oct. 7, 2011.

53118-0043 254129 Hempel, K., V. Strauss, A. Gröters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Repeated dose 28-day oral toxicity study in beagle dogs - administration in thediet,” BASF SE, Ludwigshafen, Germany, July 2, 2009. Laboratory Study # 2007/1052660. Report No. 30D0683/05077. Groups of 5 beagles/sex/group were dosed in diet with BAS 700 F(fluxapyroxad, Batch COD-000899, purity 99.7%) for 34-38 days in a pilot study to assess doselevels for the definitive subchronic study. Dietary levels were 0, 2500, 7500, and 20000 ppm. Achieved dose levels for respective male treated groups were 74, 211, and 521 mg/kg/day. Levels in females were 85, 230, and 503 mg/kg/day. No NOEL was sought nor found. Alkalinephosphatase was elevated in both sexes at all dose levels with marked dose-response. Albuminlevels were reduced consistently at all dose levels in both sexes. This study determined that20000 ppm would be excessive for the primary subchronic study (DPR Record No. 254132): thisdose level caused excessive decline in food consumption and greatly diminished uterine and


thymic weights. The dose range selected for the subchronic study was rationally chosen, basedon the present “28-day” pilot study. Useful supplementary data. Aldous, Oct. 7, 2011.

RAT

**53118-0044 254130 Kamp, H., V. Strauss, S. Groeters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - repeated dose 90-day oral toxicity study in Wistar rats; administration in the diet,”BASF SE, Ludwigshafen, Germany, Sept. 8, 2009. Laboratory Study # 2007/1005069, ReportNo. 50C0683/05064. Groups of 10 rats/sex/group were dosed in diet BAS 700 F (fluxapyroxad,Batch COD-000826, purity 99.6%) for 90 days in a subchronic study. Dietary levels were 0,100, 500, 2000, and 6000 ppm. Achieved dose levels for respective male treated groups were6.1, 31, 126, and 407 mg/kg/day. Levels for corresponding females were 7.3, 35, 144, and 424mg/kg/day. NOEL = 100 ppm, based on centrilobular hepatocellular hypertrophy and increasedrelative liver weights in both sexes. Dose levels of 500-2000 ppm appeared to elicit onlyadaptive changes. Major findings, typically with clear dose-response, comprising that full rangeincluded the above findings in both sexes, plus thyroid follicular cell hypertrophy/hyperplasia infemales, and reduced aspartate aminotransferase in males. Hepatic single cell necrosis wasobserved in 9/10 high dose males, but not in any other groups. This would indicate an excessivedose level for the subsequent chronic study. Thyroid follicular cell hypertrophy and/orhyperplasia was observed at 2000-6000 ppm in males, and at 500-6000 ppm in females,plausibly secondary to liver metabolic induction. Acceptable, with no adverse effects. Aldous,Sept. 2, 2011.

53118-0041 254127 Kamp, H., E. Fabian, V. Strauss, W. Kaufmann, and B. van Ravenzwaay,“BAS 700 F: repeated dose toxicity study in Wistar rats; administration in the diet for 4 weeks,”BASF SE, Ludwigshafen, Germany, 10/14/09. Laboratory Study # 2009/7006273. Groups of 5Wistar rats/sex/group were dosed in diet with Fluxapyroxad (BAS 700 F), Batch 32740/173,purity 99.81%, for 4 weeks in a probe subchronic study. Treatment groups of 0, 100, 500, 2000,and 6000 corresponded to achieved dose levels of 9.0, 44, 176, and 530 mg/kg/day for increasingdose groups of treated males, and of 9.4, 48, 183, and 531 mg/kg/day in females. This was thepilot study for 53118-0044 254130, above. Since this pilot reported comparable effects to thedefinitive subchronic study, no DPR worksheet is needed. Aldous, 5/10/11.

**53118-0047 25 Kaspers, U., V. Strauss, M. C. Rey Moreno, E. Fabian, and W. Mellert, “BAS 700 F - Repeateddose 28-day dermal toxicity study in Wistar rats,” BASF SE, Ludwigshafen, Germany, 10/14/09. Laboratory Study # 2009/1072489. Groups of 10 Wistar rats were dosed on clipped dorsal skinunder semi-occlusive dressing for 6 hrs/day, 5 days/week with BAS 700 F (fluxapyroxad), BatchCOD-001049, purity 99.2%, at 0, 100, 300, or 1000 mg/kg/day. Suspensions were prepared in1% aq. CMC at 4 ml/kg b.w. NOEL = 300 mg/kg/day, based on modest liver weight elevation(statistically significant in both sexes at 1000 mg/kg/day). No other effects were identified. Study is acceptable, with no adverse effects. Aldous, Oct. 7, 2011.


MICE

53118-0045 254131 Kamp, H, V. Strauss, A. Groeters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Repeated dose 90-day oral toxicity study in C57BL/6 J Rj mice; administration inthe diet,” BASF SE, Ludwigshafen, Germany, Sept. 11, 2009, Laboratory Study #2007/1018641. Report No. 51C0683/05070. Groups of ten C57BL/6 J Rj mice/sex/group weredosed in diet BAS 700 F (fluxapyroxad, Batch COD-000826, purity 99.6%) for 92-93 days in asubchronic pilot study at 0, 100, 400, 2000, and 6000 ppm. Mean achieved dose levels were 21,77, 390, and 1136 mg/kg/day for increasing dose levels in treated males, and 32, 128, 610, and1657 mg/kg/day in females. NOEL = 100 ppm for males, based on decreased cholesterol andtriglycerides, and possibly treatment-related centrilobular fatty change. NOEL for females = 400ppm, based on decreased albumin and decreased cholesterol. Liver weights were elevated at2000 and 6000 ppm in both sexes. The highest dose level caused reduced food consumption inboth sexes and reduced body weight in males. Study is supplementary by design, undertaken toset dose levels for the mouse oncogenicity study. Aldous, 5/31/11.

53118-0042 254128 Kamp, H, V. Strauss, A. Groeters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Repeated dose toxicity study in C57BL/6 J Rj mice; administration in the diet for4 weeks,” BASF SE, Ludwigshafen, Germany, Sept. 11, 2009. Laboratory Study #2007/1005068. Report No. 31C0683/05067. This was the pilot study which helped set doselevels for the subchronic mouse study, Record No. 254131, above. This pilot study has nobearing on NOEL’s or toxicity characterization. No DPR worksheet. Aldous, June 1, 2011.

MECHANISTIC STUDIES RELATING TO COMBINED, RAT

53118-0061 254165 Buesen, R., V. Strauss, E. Fabian, S. Groeters, and B. van Ravenzwaay,“BAS 700 F: Enzyme induction in liver of Wistar rats - administration in the diet over 2 weeksand recovery period of about 4 weeks,” BASF SE, Ludwigshafen, Germany, 10/28/09. BASFRegistration Document No. 2009/1072495. Ten Wistar rats/sex/group were dosed withfluxapyroxad (99.2% purity) in diet for 2 weeks at 0, 250, 1500, or 3000 ppm. Achieved doselevels in treated males were 16, 96, and 192 mg/kg/day, and in females: 19, 126, and 234mg/kg/day. Additional groups of 10/sex were similarly dosed for 2 weeks at 0 or 3000 ppm,then taken off treatment for 4 weeks to assess recovery. At term of treatment or recovery, ratswere necropsied and examined for histopathology. In addition, liver microsomes were assayedfor total cytochrome P-450 content, for activities of primary metabolism: ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-depentylase (PROD), and benzoxyresorufin-O-debenzylase (BROD); and of glucuronosyltransferase activities MUF-GT and HOBI-GT (for 4-methylumbelliferone and 4-hydroxybiphenyl substrates), and the T4-specific UDP-glucuronosyltransferase. Liver weights were significantly elevated in all male groups (107%,135%, and 152% of control weights in increasing dose groups) and in 1500-3000 ppm females(120% and 144% of controls, respectively). Liver weights were near normal in recovery rats. Thyroid gland weights were significantly elevated in 1500 ppm males (120% of controls),indicating treatment effect. Of thyroid-associated hormones, TSH was significantly elevated in3000 ppm males only (150% at termination). T3 and T4 levels were unaffected. There were noresidual effects on these hormones after recovery. Thyroid follicular hypertrophy/hyperplasiawas observed in 250-3000 ppm males and in high dose females. All high dose rats had liver


centrilobular hypertrophy, as also most rats at 1500 ppm rats, and some at 250 ppm. No suchhistopathology persisted in either organ in recovery rats. Cytochrome P-450 content (ng/mgprotein basis) on day 15 was elevated significantly in all groups, to a maximum extent of about2-fold. EROD activities peaked at about 3-fold over controls. PROD activities were induced upto 20-fold in males and up to 125-fold in females. BROD activities were induced up to 10-foldin males and 127-fold in females. Activity of MUF-GT was increased up to 5-fold in males and4-fold in females. Similarly, HOBI-GT was elevated up to 5-fold in males and 3-fold in females. T4-UDP-glucuronosyltransferase activity was increased up to a comparatively modest 1.6-foldfor males and 2.7-fold for females. The latter is evidently a key factor in rodent thyroidfollicular cell cancer induction. In contrast, the primary means of inactivation of T4 in humansis deiodination (A. Parkinson, p. 167, in Klaassen, C.D., Ed., Casarett and Doull’s Toxicology:The Basic Science of Poisons, Fifth Edition, New York, McGraw-Hill, 1996). Treatment-relatedincreases in the above metabolic enzyme activities were largely or completely reversed inrecovery rats. Useful supplementary data. Aldous, Sept. 1, 2011.

53118-0060 254164 Buesen, R., E. Fabian, S. Groeters, and B. van Ravenzwaay, “BAS 700 F:Enzyme induction in liver of Wistar rats - administration in the diet over 2 weeks,” BASF SE,Ludwigshafen, Germany, 2/25/10, Laboratory Study # 2009/1072496. Ten Wistar rats/sex/groupwere dosed with fluxapyroxad (99.2% purity) in diet for 2 weeks at 0 or 50 ppm (3.0 and 3.8mg/kg/day in males and females, respectively). This is a follow-up of BASF RegistrationDocument # 2009/1072495 (DPR Record No. 254165), which identified liver enzyme inductiondown to the lowest dose tested (250 ppm). At termination of the present study, rats werenecropsied, and liver microsomes were assayed for total cytochrome P-450 content, and for thefollowing liver microsomal activities: ethoxyresorufin-O-deethylase (EROD), pentoxyresorufin-O-depentylase (PROD), and benzoxyresorufin-O-debenzylase (BROD); and ofglucuronosyltransferase activities MUF-GT and HOBI-GT (for 4-methylumbelliferone and 4-hydroxybiphenyl substrates), and the T4-specific UDP-glucuronosyltransferase. Treatmentcaused a statistically significant increase in BROD activity in both sexes (134% of controls inmales, and 421% of controls in females). Also significant was increased HOBI-GT activity inmales (123% of controls). Useful supplementary information, extending the dose-responserelationships to activities below levels of statistical significance in most cases, or to levels whichallow extrapolation for well-defined response curves. Aldous, Oct. 6, 2011.

53118-0065 254170 Buesen, R., V. Strauss, E. Fabian, S. Groeters, and B. van Ravenzwaay,“BAS 700 F - Thyroid hormone study: repeated dose oral toxicity study in Wistar rats -administration via the diet for 4 weeks,” BASF SE, Ludwigshafen, Germany, 10/20/09. Laboratory Study # 2009/1072497. Ten Wistar rats/sex/group were dosed with fluxapyroxad(99.2% purity) in diet for 4 weeks at 0, 50, 250, 1500, or 3000 ppm. Mean achieved dose levelswere 3.5, 19, 105, and 214 mg/kg/day in increasing treated male groups, and 4.4, 20, 117, and237 mg/kg/day in females. Investigators sampled blood of fasted rats pre-test and on treatmentdays 3, 7, 14, 21, and 28 to assess thyroid-related hormones: T3, T4, and TSH. Investigatorsrecorded liver and thyroid weights and gross changes, but did not examine tissuesmicroscopically. Absolute NOEL = 50 ppm in males and 250 ppm for females, based onelevated liver weights. NOEL for thyroid-associated hormone levels (parameters of primaryinterest in this study) = 1500 ppm, based on measurably reduced T4 and elevated TSH in 3000ppm males only. T3 was unaffected. Liver weights were elevated to 133% and 150% ofcontrols in 1500 ppm and 3000 ppm males, and to 122% and 149% in corresponding females. Aldous, April 1, 2011.


53118-0062 254166 Buesen, R., W. Kaufmann, E. Fabian, and B. van Ravenzwaay, “BAS 700F - S-phase response study in Wistar rats: administration via the diet for 1, 3, 7 and 14 days,”BASF SE, Ludwigshafen, Germany, 2/25/10. Laboratory Study # 2009/1072500. Ten Wistarrats/sex/group were dosed in diet with BAS 700 F [Fluxapyroxad], Batch COD-001049, purity99.2%, for 1, 3, 7, or 14 days in a supplementary study at 0, 50, 250, 1500, or 3000 ppm. Representative (7-day cohort) achieved dose levels were 3.3, 16, 100, and 183 mg/kg/day fortreated males, and 3.5, 17, 92, and 195 mg/kg/day for females. Key parameters examined wereliver weights, liver histopathology, and particularly liver cell proliferation (7 days afterimplanting osmotic pumps delivering BrdU). Liver weights of both sexes were consistentlyelevated at 1500 and 3000 ppm following treatment of 3 to 14 days. A slight increase in liverweight at day 14 in 250 ppm males was also likely treatment-related. Livers showed dose-related increases in incidence and degree of hepatocellular hypertrophy in males at 250 ppm andabove, and in females at 1500 to 3000 ppm. Hypertrophy was observed on days 7-14 in males. Hypertrophy was observed in all 1500-3000 ppm females at day 14, but only in 3000 ppmfemales during days 3-7. Hepatic cell proliferation was increased after 3-day and 7-daytreatments in 1500 and 3000 ppm males and females in all zones. In males the strongestresponse was in zone 2 (19 to 30-fold), and secondarily in zone 3 (9 to 19-fold) on days 3 and 7. At day 14, the only significant increases in males were in zone 3 (3-fold at 1500 and 3000 ppm). In females, the strongest response was clearly zone 3 (centrilobular). Proliferation wassignificantly increased in 250 ppm females, primarily in zone 3, at days 3-14 (7 to 17-fold). Proliferation in 1500-3000 ppm females was strong, consistent, and dose-related. A transientincrease was observed at 50 ppm in day 7 females only. Investigators concluded that test articlewas behaving in the manner of an enzyme inducer such as phenobarbital. NOEL in males = 250ppm for hepatocellular cell proliferation, and 50 ppm for hepatocellular hypertrophy and liverweight increases. NOEL in females is slightly below 50 ppm for hepatocellular cellproliferation, and 250 ppm for hepatocellular hypertrophy. Males (1500-3000 ppm, treatmenttimes of 7-14 days) had slightly elevated thyroid weights, but NOEL could not be framedexactly. Useful supplementary information, exploring shorter exposure time frames (mostrelevant times for proliferation) and a lower extension of dose range than Record No. 254167. Aldous, 8/31/11.

53118-0063 254167 Buesen, R., W. Kaufmann, E. Fabian, and B. van Ravenzwaay, “BAS 700F - S-phase response study in Wistar rats: administration in the diet for 7, 28, and 91 days,”BASF SE, Ludwigshafen, Germany, 2/25/10. Laboratory Study # 2009/1072498. Groups of 10Wistar rats/sex/group were dosed in diet with BAS 700 F [Fluxapyroxad], Batch COD-001049,purity 99.2%, for 7, 28, or 91 days in a supplementary study at 0, 250, 1500, or 3000 ppm. Achieved dose (for 91-day groups) were 0, 13, 80, and 163 mg/kg/day for males, and 0, 17, 106,and 190 mg/kg/day for females. An additional 3000 ppm group was treated for 28 days,followed by a 28-day recovery period. Key parameters examined were liver weights, liverhistopathology, and liver cell proliferation (7 days after implanting osmotic pumps deliveringBrdU). Liver weights of both sexes were elevated at 1500 and 3000 ppm (statistically significantin all cases for relative liver weights) following 7, 28, and 91 days. High dose male liverweights were comparable to control after 28-day recovery, whereas high dose female relativeliver weights were modestly but significantly elevated (11% over controls after recovery,compared to 45% over controls at day 91). Livers showed dose-related increases in incidenceand degree of hepatocellular hypertrophy in all dose groups at all treatment times, but no residualeffects in recovery rats. Hepatic cell proliferation was increased after 7 days in 1500 and 3000ppm males in all zones, with the strongest response in zone 3 (centrilobular): 14- and 21-fold for


1500 and 3000 ppm, respectively. Proliferation was also increased 3-fold in 3000 ppm males inzone 3 after 91 days. In contrast, centrilobular proliferation was greatly increased in all femalegroups at all treatment intervals, with high dose increases of 26-fold, 14-fold, and 7-fold infemales at 7, 28, and 91 days, respectively. Low dose female zone 3 responses were 4-fold, 7-fold, and 3-fold at 7, 28, and 91 days, respectively. Proliferation was not increased over controlsin recovery male or female groups. Investigators concluded that test article was behaving in themanner of an enzyme inducer such as phenobarbital. Useful supplementary information. Aldous, 8/31/11.

53118-0064 254168 Buesen, R., W. Kaufmann, E. Fabian, and B. van Ravenzwaay, “BAS 700F - S-phase response study in Wistar rats: administration in the diet for 7, 28 and 91 days,”BASF SE, Ludwigshafen, Germany, 2/25/10. Laboratory Study # 2009/1072499. Ten Wistarrats/sex/group were dosed in diet with BAS 700 F [Fluxapyroxad], Batch COD-001049, purity99.2%, for 7, 28, or 91 days in a supplementary study at 0 or 50 ppm. Mean achieved doses (for91-day treated males and females, respectively) were 3.0 and 3.5 mg/kg/day. This study was afollow-up to supplementary study 53118-0063 254167, which employed dose levels of 250,1500, and 3000 ppm. That study did not identify a NOEL for centrilobular proliferation infemales, females being more responsive than males to centrilobular proliferation. There was notreatment effect in the present study at 50 ppm. There were statistical increases at 28 days infemales in liver zones 1 and 2 in this study: these are unlikely to be toxicologically relevantbecause (1) Zone 1 values for the 50 ppm females at 7 days and 91 days were lower thancontrols, and (2) the concurrent control labeling index (LI) for Zone 2 was unusually low [lessthan 50% of the LI in the main supplementary study (Record No. 254167)]. Thus an overallNOEL for centrilobular proliferation is 50 ppm for females, based on this study. The NOEL formales, based on Record No. 254167, is 250 ppm. Useful supplementary data. Aldous, 8/31/11.

53118-0066 254171 Buesen, R., E. Fabian, and B. van Ravenzwaay, “BAS 700 F - Thyroidfunction test in Wistar rats using perchlorate discharge as a diagnostic test - administration viathe diet over 2 weeks,” BASF SE, Ludwigshafen, Germany, 10/19/09. Laboratory Study #2009/1072501. Twelve rats/sex/group were dosed in diet for 2 weeks with control diet, 3000ppm fluxapyroxad, 2000 ppm propylthiouracil (PTU: which blocks “organification” of follicularcell iodine), or 1000 ppm phenobarbital sodium (PB). On day 14, all rats received 0.5 ml (1 µCi)of labeled NaI (125-I) ip. Six hrs later, 6 of these rats/sex/group received KClO4 ip (amount notspecified: function apparently to block iodide transfer across thyroid follicular cell membrane):remaining rats received saline blank. Rats were sacrificed 2.5 min after KClO4 or salinetreatment. Investigators calculated thyroid weight, specific radioactivity in blood and in thyroid,and the ratio of 125-I of thyroid to blood. Fluxapyroxad behaved similarly to PB, with thyroidweights increased up to 50% and a similar increase in specific radioactivity in thyroid. PTUelicited a 4-5-fold increase in thyroid weights, with a profound reduction of specific radioactivityin thyroid (only about 20% of control). KClO4 further reduced 125-I label in thyroids of PTU rats(to 6-11% of controls), with no effect on PB or fluxapyroxad. Results showed that fluxapyroxadbehaves similarly to PB in the outcomes assayed, and that fluxapyroxad does not inhibit thyroidhormone formation in the manner of PTU. Useful supplementary information. Aldous, 4/5/11.

53118-0064 254169 Doi, A. M., “Evaluation of a mode of action for liver tumor induction byBAS 700 F,” [not a laboratory study, but “performing facility” was BASF in Research TrianglePark, NC], 2/17/10. This author summarized key points of the above studies (mainly the rat


combined study, plus the studies in the present “Mechanistic Studies” section of this Summaryof Toxicology Data, concluding on p. 17 that “The overall weight of evidence strongly supportsa mitogenic MoA with clear thresholds for development [of] liver tumors in rats.” This is not a“study” for DPR Data Review Group to evaluate, but is a potential useful overview for liver andthyroid cancer-associated risk assessment. No DPR worksheet. Aldous, 8/29/11.

IMMUNOTOXICITY

**53118-0060 254163 Kaspers, U., V. Strauss, S. Groeters, E. Fabian, and B. van Ravenzwaay,“BAS 700 F - Immunotoxicity study in male C57BL/6 J Rj mice; administration in the diet for 4weeks,” BASF SE, Ludwigshafen, Germany, Oct. 8, 2009. Laboratory Study # 2009/1072494,Project. No. 43S0683/05105. Groups of 8 male mice/sex/group were dosed in diet withfluxapyroxad (Batch COD-001049, purity 99.2%) for 28 days at 0, 500, 2000, or 6000 ppm (0,106, 450, and 1323 mg/kg/day, respectively). Positive controls received 12 mg/kg/daycyclophosphamide (gavage) for 4 weeks. Mice were immunized with SRBC ip on day 23 toassess T-cell antibody response [by anti-SRBC (sheep blood RBC) IgM ELISA methods]. Sacrifice was on day 29. Natural killer (NK) cell activity was assessed for leukocytes derivedfrom spleens by determining damage to YAC lymphoma cells by flow cytometry after stainingfor damaged YAC cells. Flow cytometry was used to differentiate sub-populations oflymphocytes, with aid of stain-labeled antibodies to mouse CD3,, CD4, CD8a, NK-1.1, andCD19. Fluxapyroxad was negative for all parameters. Cyclophosphamide greatly reduced T-cell antibody response, markedly reduced all sub-populations of lymphocytes assayed, andsharply reduced B:T ratios. Cyclophosphamide caused a profound decrease in T-cell antibodyresponse, but no effect on NK cell activity. Acceptable, with no adverse effects. Aldous, Oct. 6,2011.

TOXICITY STUDIES ON TECHNICAL IMPURITIES (generally no DPR worksheets needed)

53118-0067 254189 Schulz, M., and Landsiedel, R., “Reg. No. 5425764 (Technical impurity ofBAS 700F) - Salmonella typhimurium / Escherichia coli reverse mutation assay (standard platetest and preincubation test),” BASF AG, Ludwigshafen, Germany, 1/18/08, Laboratory Study #2007/1054385. There were no consistent increases in revertants. A marginal (2 x) increase inthe standard plate test with TA 1535 without S-9 at 500 µg/plate was not repeated in a standardplate test nor in the pre-incubation test. Aldous, no treatment effects, and no DPR worksheet,July 6, 2011.

53118-0067 254190 Schulz, M., and Landsiedel, R., “Reg. No. 5425764 (Technical impurity ofBAS 700F) - in vitro gene mutation test in CHO cells (HPRT locus assay),” BASF SE,Ludwigshafen, Germany, Nov. 4, 2008, Laboratory Study # 2008/1068013. There were notreatment effects indicated in either of two separate experiments with or without S-9. Aldous, noDPR worksheet, July 6, 2011.

53118-0067 254191 Schulz, M., and Landsiedel, R., “Reg. No. 5425764 (Technical impurity ofBAS 700F) - micronucleus test in bone marrow cells of the mouse,” BASF AG, Ludwigshafen,Germany, 1/17/08, Laboratory Study # 2008/1002421. There were no treatment effects indicatedin either of two separate experiments, the first experiment with dose levels of 0, 500, 1000, and


2000 mg/kg Reg. No. 5425764 (purity 99.9%) with a 24-hr sacrifice interval, nor at 0 and 2000mg/kg with a 48-hr period. Aldous, no DPR worksheet, Sept. 6, 2011.

53118-0068 254192 Schulz, M., and Landsiedel, R., “Reg. No. 5356469 (Technical impurity ofBAS 700F) - Salmonella typhimurium / Escherichia coli reverse mutation assay (standard platetest and preincubation test),” BASF AG, Ludwigshafen, Germany, 1/28/08, Laboratory Study #2007/1054448. There were no remarkable, dose-related, or consistent increases in revertants. Aldous, no treatment effects, and no DPR worksheet,7/7/11.

53118-0068 254193 Schulz, M., and Landsiedel, R., “Reg. No. 5356469 (Technical impurity ofBAS 700F) - in vitro gene mutation test in CHO cells (HPRT locus assay),” BASF SE,Ludwigshafen, Germany, Nov. 4, 2008, Laboratory Study # 2008/1068014. There were notreatment effects indicated in either of two separate experiments with or without S-9. Aldous, noDPR worksheet, 7/7/11.

53118-0068 254194 Schulz, M., and Landsiedel, R., “Reg. No. 5356469 (Technical impurity ofBAS 700F) - micronucleus test in bone marrow cells of the mouse (including amendment No. 1,”BASF SE, Ludwigshafen, Germany, 12/19/08 (amended), Laboratory Study # 2008/7020156. There were no treatment effects indicated in either of two separate experiments, the firstexperiment with dose levels of 0, 500, 1000, and 2000 mg/kg fluxapyroxad with a 24-hr sacrificeinterval, nor at 0 and 2000 mg/kg with a 48-hr period. Aldous, no DPR worksheet, 7/7/11.

53118-0069 254199 Schulz, M., and Landsiedel, R., “3',4',5'-Trifluoro-biphenyl-2-ylamine:Salmonella typhimurium / Escherichia coli reverse mutation assay (standard plate test and preincubation test),” BASF SE, Ludwigshafen, Germany, Oct. 1, 2009, Laboratory Study #2009/1072516. There were no remarkable alterations in revertants. Aldous, no treatmenteffects, and no DPR worksheet,7/7/11.

53118-0069 254200 Schulz, M., and Landsiedel, R., “3',4',5'-Trifluoro-biphenyl-2-ylamine - invitro gene mutation test in CHO cells (HPRT locus assay),” BASF SE, Ludwigshafen, Germany,9/22/09, Laboratory Study # 2009/1072518. There were no treatment effects indicated in eitherof two separate experiments with or without S-9. Aldous, no DPR worksheet, 7/7/11.

53118-0069 254201 Schulz, M., and Landsiedel, R., “3',4',5'-Trifluoro-biphenyl-2-ylamine:micronucleus test in bone marrow cells of the mouse,” BASF SE, Ludwigshafen, Germany, Oct.2, 2009, Laboratory Study # 2009/1072517. There were no treatment effects indicated in eitherof two separate experiments, the first experiment with dose levels of 0, 15, 30, and 60 mg/kg of3',4',5'-trifluoro-biphenyl-2-ylamine with a 24-hr sacrifice interval, nor at 0 and 60 mg/kg with a48-hr period. Micronuclei in the 60 mg/kg group with the 48-hr period were significantlyelevated over concurrent controls (0.8 vs. 1.8 micronuclei per thousand PCE’s), but was similarto mean historical control values, and identical to a control value reported in the same month bythese investigators (DPR Record No. 254216). Toxicity was evident at all dose levels tested:with piloerection at 15 mg/kg, and additionally hunched posture and “reduced general condition”at 30-60 mg/kg, and “necrotic tail tip” in all 60 mg/kg mice at 48 hrs. Aldous, no DPRworksheet, 9/6/11.


TOXICITY STUDIES ON METABOLITES

BASF Reg. No. 5435595

NOTE: This test article is designated as M700F002 in fluxapyroxad metabolism studies. IUPACname: 3-(difluoromethyl)-1H-pyrazole-4-carboxylic acid. Purity = 99.3% (from Record No.254209, p. 14).

53118-0074 254209 Cords, S.-M., and E.-M. Lammer, “Reg. No. 5435595 (Metabolite of BAS700F) - acute oral toxicity study in rats,” Bioassay Labor Für Biologische Analytic GmbH,Heidelberg, Germany. Six female Wistar rats were dosed once by gavage at 2000 mg/kg. Norats died. Clinical signs of “impaired general state,” dyspnea, and piloerection were observed in2/6 rats. LD50 (F) > 2000 mg/kg. Toxicity Category III. Useful data, no DPR worksheet,Aldous, 8/24/11.

53118-0074 254210 Kaspers, U., V. Strauss, S. Gröters, E. Fabian, and B. van Ravenzwaay,“Reg. No. 5435595 (Metabolite of BAS 700F) - Repeated dose 28-day oral toxicity study inWistar rats; administration in the diet,” BASF SE, Ludwigshafen, Germany, Dec. 8, 2008. FiveWistar rats/sex/dose were dosed in diet at 0, 1500, 5000, or 15000 ppm for 4 weeks. There wereno treatment-related clinical signs, no changes in hematology or clinical chemistry, nor changesin necropsy or histopathology. Thus the NOEL is 15000 ppm, estimated to be 1165 mg/kg/dayfor males, and 1253 for females. Useful data, no DPR worksheet, Aldous, 8/24/11.

53118-0075 254211 Kaspers, U., V. Strauss, S. Gröters, E. Fabian, and W. Mellert, “Reg. No.5435595 (Metabolite of BAS 700F) - Repeated dose 90-day oral toxicity study in Wistar rats;administration in the diet,” BASF SE, Ludwigshafen, Germany, 5/13/09. Dietary levels wereadjusted weekly to achieve target dose levels of 0, 100, 300, or 1000 mg/kg/day. There were 10rats/sex/group. Investigators assessed clinical signs, and at or near to termination, alsohematology, clinical chemistry, urinalysis, ophthalmology, necropsy, and histopathology. Notreatment-related effects were observed. NOEL = 1000 mg/kg/day for both sexes. Useful data,no DPR worksheet, Aldous, 8/24/11.

53118-0076 254212 Schneider, S., E. Fabian, and W. Mellert, “Reg. No. 5435595 (Metaboliteof BAS 700F) - Prenatal developmental toxicity study in New Zealand White rabbits - oraladministration (gavage),” BASF SE, Ludwigshafen, Germany, Oct. 13, 2009. Rabbits receivedgavage treatments of 0, 100, 300, and 1000 mg/kg/day from gestation days 6-28 in a standarddevelopmental toxicity study. Incidences of abortions, and premature deaths were elevated at1000 mg/kg/day. Significantly reduced food consumption, absent or reduced defecation, andoccasional stomach ulceration at necropsy were major indicators of poor general state at the highdose. Body weight gain in surviving high dose does was significantly reduced. Developmentaltoxicity was not evident at any dose. Maternal NOEL = 300 mg/kg/day (mortalities and clinicalsigns). Developmental NOEL = 1000 mg/kg/day (no treatment-related effects at HDT). Usefuldata. No DPR worksheet is warranted at this time. Aldous, 10/10/11.

53118-0077 254213 Schulz, M., and Landsiedel, R., “Reg. No. 5435595 (Metabolite of BAS700F) - Salmonella typhimurium / Escherichia coli reverse mutation assay (standard plate testand preincubation test),” BASF AG, Ludwigshafen, Germany, 11/21/07, Laboratory Study #2007/1051931. There were no increases in revertants. Aldous, no DPR worksheet,7/7/11.


53118-0077 254214 Schulz, M., and Landsiedel, R., “Reg. No. 5435595 (Metabolite of BAS700F) - in vitro gene mutation test in CHO cells (HPRT locus assay),” BASF SE, Ludwigshafen,Germany, 7/14/08, Laboratory Study # 2008/1014199. There were no treatment effects indicatedin either of two separate experiments without S-9 or three separate experiments with S-9. Aldous, no DPR worksheet, 7/7/11.

53118-0077 254215 Schulz, M., and Landsiedel, R., “Reg. No. 5435595 (Metabolite of BAS700F) - in vitro chromosomal aberration assay in V79 cells,” BASF SE, Ludwigshafen,Germany, March 10, 2008, Laboratory Study # 2008/1002741. Dose levels from 100 to 1600µg/ml were employed, with hours of exposure/preparation period without S-9 of 4/18 (twoexperiments), 18/18, and 18/28. There were no significant elevations without S-9 due to thetested metabolite, although 800-1600 µg/ml groups exceeded reported historical control values. Concurrent control was relatively high. The same dose levels were used with S-9, with hours ofexposure/preparation period of 4/18 and 4/28. There were statistically significant elevationswith S-9 in metabolite groups, but these were within reported historical control values, lackeddose-response, and appear to be negative. Investigators justifiably considered the study asnegative. Aldous, no DPR worksheet, 7/7/11.

53118-0077 254216 Schulz, M., and Landsiedel, R., “Reg. No. 5435595 (Metabolite of BAS700F): micronucleus test in bone marrow cells of the mouse ,” BASF SE, Ludwigshafen,Germany, Oct. 5, 2009. Laboratory Study # 2009/1072508. Dose levels of 0, 375, 750, and1500 mg/kg Reg. No. 5435595 (99.3% purity) were administered to 5 NMRI mice/group, 24 hrsbefore sacrifice. Additional groups at 0 and 1500 mg/kg were evaluated 48 hrs after dosing. There were no clinical signs in this range, however 2000 mg/kg had been shown to elicit majorclinical signs for at least 48 hrs, hence maximum dose for final study was justified. There wereno treatment-related increases in micronuclei. Useful data, no DPR worksheet, Aldous, 9/6/11.

53118-0077 254217 Fabian, E., and Landsiedel, R., “Reg. No. 5435595 (Metabolite of BAS700F): study on the kinetics in mice, BASF SE, Ludwigshafen, Germany, Oct. 9, 2009. Laboratory Study # 2009/1098042. This study related to the mouse micronucleus study, andconfirmed that a high dose (1000 mg/kg) reached mean concentrations in bone marrow at 5 hrswhich were about double that of blood cells, with plasma residues intermediate between bloodcells and marrow. Thus a meaningful dose was received in the marrow for a valid micronucleusstudy. Useful data, no DPR worksheet, Aldous, 7/7/11.


NOTE: This test article is a very minor metabolite in the rat. It is CAS No. 176969-34-9:1H-Pyrazole-4-carboxylic acid, 3-(difluoromethyl)-1-methyl- [99.2% purity] (see Document No.53118-0072, Record No. 254204, p. 551. This substance is designated as M700F001 in themetabolism studies of this Summary.

53118-0070 254202 Cords, S.-M., and E.-M. Lammer, “Reg. No. 5069089 (Metabolite of BAS700F): acute oral toxicity study in rats,” Bioassay Labor Fuer Biologische Analytic GmbH,Heidelberg, Germany, Laboratory Study No. 2009/1072502. Six female Wistar rats were dosedonce with Reg. No. 5069089 (purity 99.2%) by gavage at 2000 mg/kg. One rat had generalclinical signs during hours 4-5 after dosing: “impaired general state,” dyspnea, and piloerection.


Reduced feces were observed in 2 rats. There were no deaths. LD50 (F) > 2000 mg/kg. ToxicityCategory III. Useful data, no DPR worksheet, Aldous, 9/6/11.

53118-0071 254203 Kaspers, U., V. Strauss, S. Groeters, E. Fabian, and B. van Ravenzwaay, “ Reg. No. 5069089 (Metabolite of BAS 700F) - Repeated dose 90-day oral toxicity study inWistar rats; administration in the diet,” BASF SE, Ludwigshafen, Germany, Oct. 8, 2009. Report No. 50S0451/07119. (Reg. No. 5069089, Batch No. L80-68, purity 99.2%). This studyindicated no remarkable findings up to a dose level of 1000 mg/kg/day in the report summary. No DPR worksheet is warranted at this time. Aldous, 8/23/11.

53118-0072 254204 Schneider, S., E. Fabian, and W. Mellert, “Reg. No. 5069089 (Metaboliteof BAS 700F) - Prenatal developmental toxicity study in New Zealand White rabbits - oraladministration (gavage),” BASF SE, Ludwigshafen, Germany, Oct. 14, 2009. Report No.40R0451/07118. Reg. No. 5069089 (Batch No. L80-68, purity 99.2%). This study indicated noremarkable findings in the report summary up to the highest dose tested (250 mg/kg/day). Someclinical signs reported in the associated pilot study included reduced food consumption at 250mg/kg/day, and occasional stomach erosions, and mortalities and/or abortion at 500 to 1000mg/kg/day. Tables of fetal data from the primary study showed no treatment-related findings. No DPR worksheet is warranted at this time. Aldous, 8/25/11.

53118-0073 254205 Schulz, M. and R. Landsiedel, “Reg. No. 5069089 (Metabolite of BAS700F) - Salmonella typhimurium / Escherichia coli reverse mutation assay (standard plate testand preincubation test),” BASF SE, Ludwigshafen, Germany, Sept. 7, 2009. Reg. No. 5069089 (Batch No. L80-68, purity 99.2%). Report No. 40R0451/074195. This test was negative,employing concentrations up to 5000 µg/plate (or 2x or 4x reduction thereof if limited bycytotoxicity). No DPR worksheet is warranted at this time. Aldous, 8/24/11.

53118-0073 254206 Schulz, M. and R. Landsiedel, “Reg. No. 5069089 (Metabolite of BAS700F) - in vitro gene mutation test in CHO cells (HPRT locus assay) (Including AmendmentNo. 1),” BASF SE, Ludwigshafen, Germany, July 9, 2009, Amended Sept. 1, 2009. Report No.50M0451/074157. Reg. No. 5069089 (Batch No. L80-68, purity 99.2%). Dose levels up to2000 µg/mL (about 11.4 mM, meeting limit test) were not limited by toxicity. Primary test used250, 500, 1000, and 2000 µg/mL with 4 hr or 24 hr exposure periods, each with and without S-9. Results were negative. No DPR worksheet is warranted at this time. Aldous, 8/24/11.

53118-0073 254207 Schulz, M. and R. Landsiedel, “Reg. No. 5069089 (Metabolite of BAS700F) - in vitro chromosome aberration assay in V79 cells,” BASF SE, Ludwigshafen, Germany,8/31/09. Report No. 32M0451/074158. Dose levels up to 2000 µg/mL of Reg. No. 5069089(Batch No. L80-68, purity 99.2%) were employed for exposure/(time to harvest) periods of 4/18,18/18, or 18/28 hrs without S-9, and 4/18, and 4/28 hrs with S-9. All test article groups werenegative. No DPR worksheet is warranted at this time. Aldous, 8/24/11.

53118-0073 254208 Schulz, M. and R. Landsiedel, “Reg. No. 5069089 (Metabolite of BAS700F) - micronucleus test in bone marrow cells of the mouse ,” BASF SE, Ludwigshafen,Germany, Oct. 5, 2009. Report No. 26M0451/074181. Investigators tested 500, 1000, or 2000mg/kg of Reg. No. 5069089 (Batch No. L80-68, purity 99.2%) to 5 male NMRI mice/group witha 24-hr sacrifice time, or 2000 mg/kg with a 48-hr sacrifice time. This treatment did not increasemicronuclei. No DPR worksheet is warranted at this time. Aldous, 8/24/11.



NOTE: This test article was characterized in the report as a metabolite of fluxapyroxad,(designated M700F007 in the metabolism studies) purity 99.4%.

53118-0078 254218 Cords, S.-M., and E.-M. Lammer, “Reg. No. 5621781 (Metabolite of BAS700F): acute oral toxicity study in rats,” Bioassay Labor Fuer Biologische Analytic GmbH,Heidelberg, Germany, Laboratory Study No. 2009/1084176, Oct. 2, 2009. Three female Wistarrats were dosed once at 2000 mg/kg of Reg. No. 5621781, Batch L81-108, purity 99.4%. All ofthese died: 2 of them within 5 hours after dosing. All had multiple clinical signs such as “poorgeneral state,” dyspnea, ataxia, twitching, staggering, and piloerection. Six females were dosedonce with 500 mg/kg. Of these, one showed some of the above signs during hrs 4-5: all otherswere without unusual clinical signs. Necropsy was not remarkable in any group. Thus 500mg/kg < LD50 (F) < 2000 mg/kg. Toxicity Category III. Useful data, no DPR worksheet,Aldous, 8/25/11.


NOTE: This test article was characterized in Document No. 53118-0083, Record No. 254227,(page 29) as M700F048. It is the $-glucuronide of fluxapyroxad, the glucuronide displacing theN-methyl group of an otherwise intact fluxapyroxad molecule. Most of the glucuronides derivedfrom the a.i. also had hydroxylation on one of the phenyl rings (Document No. 53118-0058, Record No. 254161) so that this is not one of the most abundant metabolites.

53118-0079 254219 Cords, S.-M., and E.-M. Lammer, “Reg. No. 5570265 (Metabolite of BAS700F): acute oral toxicity study in rats,” Bioassay Labor Fuer Biologische Analytic GmbH,Heidelberg, Germany, Laboratory Study No. 2009/1018496, 7/14/09. Six female Wistar ratswere dosed once at 2000 mg/kg of Reg. No. 5570265, Batch No. L81-104, purity 93.7%. Nodeaths occurred. Necropsy was not remarkable in any group. Some clinical signs were observedin 4 rats, primarily within the first 3 hrs or up to a maximum of 3 days. Signs included diarrhea,impaired general state, piloerection, and dyspnea. LD50 (F) > 2000 mg/kg. Toxicity CategoryIII. Useful data, no DPR worksheet, Aldous, 8/25/11.

53118-0080 254220 Kaspers, U., V. Strauss, S. Groeters, E. Fabian, and B. van Ravenzwaay,“Reg. No. 5570265 (Metabolite of BAS 700F) - Repeated dose 28-day oral toxicity study inWistar rats; administration in the diet,” BASF SE, Ludwigshafen, Germany, Oct. 8, 2009. Laboratory Study # 2009/1072510. Ten Wistar rats/sex/dose were dosed in diet with Reg. No.5570265 (93.7% purity) for 28 days at nominal 0, 50, 200, or 1000 mg/kg/day. High dosefindings, normally in both sexes, included reduced body weight gains, elevated liver weights(absolute and relative), reduction of bile acids in serum, and increase in urinary crystals(significant in females only). In the histopathological examination, 8 males and 4 females at1000 mg/kg/day demonstrated minimal hepatocellular centrilobular hypertrophy in the liver. Inaddition, there was a dose-related reduction in total bilirubin at 200 to 1000 mg/kg/day in bothsexes. Useful data, no DPR worksheet, Aldous, 9/6/11.


53118-0081 254221 Schneider, S., E. Fabian, and W. Mellert, “Reg. No. 5570265 (Metaboliteof BAS 700F) - Prenatal developmental toxicity study in New Zealand White rabbits - oraladministration (gavage),” BASF SE, Ludwigshafen, Germany, 2/22/10. Report No. 40R0008/09008. Dose levels were 0, 10, 30, and 100 mg/kg/day of metabolite Reg. No. 5570265 (BatchL81-124, purity 96.1%), administered by gavage on gestation days 6-28. Maternal toxicity at100 mg/kg/day was reflected by decreased food consumption and decreased body weight gainduring the treatment period, and abortions usually tied to clinical signs of “no defecation.” There were no definitive fetal effects. No DPR worksheet is warranted at this time. Aldous,8/25/11.

53118-0082 254222 Schulz, M. and R. Landsiedel, “Reg. No. 5570265 (Metabolite of BAS700F) - Salmonella typhimurium / Escherichia coli reverse mutation assay (standard plate testand preincubation test),” BASF SE, Ludwigshafen, Germany, Sept. 21, 2009. Report No.40M0008/094079. This test was negative, employing concentrations in 2x steps, not limited bysolubility and generally not by toxicity up to 5500 µg/plate of Reg. No. 5570265 (Batch L81-124, purity 93.7%). No DPR worksheet is warranted at this time. Aldous, 8/25/11.

53118-0082 254223 Schulz, M. and R. Landsiedel, “Reg. No. 5570265 (Metabolite of BAS700F) - in vitro gene mutation test in CHO cells (HPRT locus assay),” BASF SE,Ludwigshafen, Germany, 8/31/09. Report No. 50M0008/094054. Dose levels up to 500-750µg/mL of Reg. No. 5570265 (Batch No. L81-104, purity 93.7%) were usable without S-9, and upto 1000-1250 µg/mL with S-9, limited by cytotoxicity. Test article was not mutagenic. No DPRworksheet is warranted at this time. Aldous, 8/25/11.

53118-0082 254224 Schulz, M. and R. Landsiedel, “Reg. No. 5570265 (Metabolite of BAS700F) - in vitro chromosomal aberration assay in V79 cells,” BASF SE, Ludwigshafen,Germany, Oct. 2, 2009. Report No. 32M0008/094198. Exposure/(time to harvest) periods of4/18, 18/18, or 18/28 hrs were used without S-9, and 4/18, and 4/28 hrs with S-9. Dose levels upto 375 to 750 µg/mL of Reg. No. 5570265 (Batch No. L81-124, purity 96.1%) were maximumusable levels without S-9, and dose levels up to 1000 to 1200 µg/mL were usable with S-9, basedon cytotoxicity. All test article groups without S-9 were negative. The top readable dose levelswith S-9 (1000 to 1200 µg/mL) elicited statistically increased chromosomal aberrations in twotrials at 4/28 [Exposure/(time to harvest)] hrs only. [There were no increases at the next lowerdose levels of 750 to 800 µg/mL in those 2 trials]. Result is positive at limits based oncytotoxicity. No DPR worksheet is warranted at this time. Aldous, 8/25/11.

53118-0082 254225 Schulz, M. and R. Landsiedel, “Reg. No. 5570265 (Metabolite of BAS700F) - micronucleus test in bone marrow cells of the mouse ,” BASF SE, Ludwigshafen,Germany, Oct. 5, 2009. Report No. 26M0008094005. Investigators tested 500, 1000, or 2000mg/kg of Reg. No. 5570265 (Batch No. L81-124, purity 96.1%) to 5 male NMRI mice/groupwith a 24-hr sacrifice time, or 2000 mg/kg with a 48-hr sacrifice time. There was a statisticallysignificant increase in micronuclei for the 1000 mg/kg mice with a 24-hr sacrifice time: all othertest groups were negative. Investigators considered the study to be negative, considering lack ofdose-response, and noting that the significant value was well within historical range. No DPRworksheet is warranted at this time. Aldous, 8/25/11.

53118-0083 254226 Schulz, M. and R. Landsiedel, “Reg. No. 5570265 (Metabolite of BAS700F): in vivo unscheduled DNA synthesis (UDS) assay in rat hepatocytes,” BASF AG,


Ludwigshafen, Germany, 10/21/09 (as amended). Report No. 80M0008/094199. LaboratoryStudy # 2009/7006262. Groups of 3 male Wistar rats/group were dosed by gavage with 0, 1000,or 2000 mg/kg of Reg. No. 5570265, purity 96.1%, (Batch No. L81-124), or by gavage with 50mg/kg 2-acetylaminofluorene (positive control) with sampling times of 3 hrs or 14 hrs tosacrifice. Net nuclear grain counts were unaffected by test article (negative for UDS). Positivecontrol was functional. Useful data. No DPR worksheet is warranted at this time. Aldous,8/26/11.

53118-0083 254227 Fabian, E., and Landsiedel, R., “Reg. No. 5570265 (Metabolite of BAS700F): study on the kinetics in mice, BASF SE, Ludwigshafen, Germany, Oct. 9, 2009. Laboratory Study # 2009/1098044. This study related to the mouse micronucleus study inRecord No. 254225 (above), and confirmed that a high dose (1000 mg/kg) of Reg. No. 5570265(Batch No. L81-104, purity 93.7%) reached mean concentrations in bone marrow at 5 hrs whichwere over twice that of blood cells, and comparable to plasma levels. Thus the dose received inthe marrow was suitable for the micronucleus study. Useful data, no DPR worksheet, Aldous,8/26/11.

53118-0083 254228 Griesser, M., “Excretion and metabolism of 14C-M700F048 after oraladministration in rats,” BASF SE, Limburgerhof, Germany, 9/22/09. Report No. 366577. Thistest article is Reg. No. 5570265, Batch No. L81-104, purity 93.7%. Four Wistar rats/sex weredosed once by gavage with 7.5 mg/kg of 14C-M700F048 (Batch No. 957-1019, radiochemicalpurity 98.3%) with label in phenyl group adjacent to amide. Excreta were evaluated for 7 days. Urine comprised 2.4% and 6.8% of administered label in males and females, respectively. Maincomponents in the urine were characterized, and were (highest component first): (1) M700F050(cleavage of N-glucuronide of Reg. No. 5570265 followed by O-glucuronidation somewhere onone of the phenyl rings), (2) M700F009 (cleavage of N-glucuronide of Reg. No. 5570265followed by hydroxylation somewhere on one of the phenyl rings) - this was a major metaboliteof fluxapyroxad, and (3) small but measurable amounts of Reg. No. 5570265. In both sexes, 26-30% of administered dose in fecal residues was comprised of administered Reg. No. 5570265,and about equal amounts were M700F009. The other common metabolite (15-16% ofadministered dose) was M700F008 (the simple cleavage of N-glucuronide from Reg. No.5570265). Findings were consistent with behavior of fluxapyroxad in the main metabolismstudy (Record No. 254161). Useful supplementary data. No DPR worksheet. Aldous, 8/26/11.

Summary of Toxicology Data - · PDF fileSUMMARY OF TOXICOLOGY DATA FLUXAPYROXAD ... (DPN) # 53118 SB 950 # (Not applicable) Original ... purity 99.7%, for 2 yrs in an oncogenicity

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